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Generation of trophoblast stem cells from rabbit embryonic stem cells with BMP4.

Tan T, Tang X, Zhang J, Niu Y, Chen H, Li B, Wei Q, Ji W - PLoS ONE (2011)

Bottom Line: Trophoblast stem (TS) cells are ideal models to investigate trophectoderm differentiation and placental development.Rabbit ES cells generated in our laboratory were induced to differentiate in the presence of BMP4 and TS-like cell colonies were isolated and expanded.These cells expressed the molecular markers of mouse TS cells, were able to invade, give rise to derivatives of TS cells, and chimerize placental tissues when injected into blastocysts.

View Article: PubMed Central - PubMed

Affiliation: Kunming Primate Research Center, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan, China.

ABSTRACT
Trophoblast stem (TS) cells are ideal models to investigate trophectoderm differentiation and placental development. Herein, we describe the derivation of rabbit trophoblast stem cells from embryonic stem (ES) cells. Rabbit ES cells generated in our laboratory were induced to differentiate in the presence of BMP4 and TS-like cell colonies were isolated and expanded. These cells expressed the molecular markers of mouse TS cells, were able to invade, give rise to derivatives of TS cells, and chimerize placental tissues when injected into blastocysts. The rabbit TS-like cells maintained self-renewal in culture medium with serum but without growth factors or feeder cells, whilst their proliferation and identity were compromised by inhibitors of FGFs and TGF-β receptors. Taken together, our study demonstrated the derivation of rabbit TS cells and suggested the essential roles of FGF and TGF-β signalings in maintenance of rabbit TS cell self-renewal.

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Rabbit TS-like cells can chimerize placentae in vivo.(A) GFP transgenic TS-like cells. (B) Whole-mount phase-contrast photographs of the undersides of chimeric placenta. White rectangle represents the part of placenta magnified in (C). (C) Magnification of part of the chimeric placenta in (B). (D) Fluorescent photograph of (C). The placenta showed an extensive contribution of GFP+ cells; Note that the blood vessels of the placenta had no fluorescence (arrow). (E) Whole-mount phase-contrast photographs of the undersides of control placenta developed from un-injected blastocyst. White rectangle represents the part of placenta magnified in (F). (F) Magnification of part of a non-chimeric placenta in (E). (G) Fluorescent photograph of (F). The control placenta showed weak background fluorescence. (H) Phase-contrast photograph of the Yolk sac from the chimeric embryo. (I) Fluorescent photograph of (H) Magnification of C, I, J is 4×, and F is 2×; Magnification of D, E is 7×, and G and H is 4×.
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pone-0017124-g004: Rabbit TS-like cells can chimerize placentae in vivo.(A) GFP transgenic TS-like cells. (B) Whole-mount phase-contrast photographs of the undersides of chimeric placenta. White rectangle represents the part of placenta magnified in (C). (C) Magnification of part of the chimeric placenta in (B). (D) Fluorescent photograph of (C). The placenta showed an extensive contribution of GFP+ cells; Note that the blood vessels of the placenta had no fluorescence (arrow). (E) Whole-mount phase-contrast photographs of the undersides of control placenta developed from un-injected blastocyst. White rectangle represents the part of placenta magnified in (F). (F) Magnification of part of a non-chimeric placenta in (E). (G) Fluorescent photograph of (F). The control placenta showed weak background fluorescence. (H) Phase-contrast photograph of the Yolk sac from the chimeric embryo. (I) Fluorescent photograph of (H) Magnification of C, I, J is 4×, and F is 2×; Magnification of D, E is 7×, and G and H is 4×.

Mentions: The above evidences prompted us to take the strictest test as if these TS-like cells were able to chimerize the placental tissues when injected into blastocysts. GFP transgenic TS-like cells (Figure 4A) were injected into blastocysts to examine their ability to chimerize the placental tissues. At day 20 of gestation, placenta proper, but not embryo proper of the conceptus (4 out of 12) derived from the green TS like-cell injected blastocysts emitted intensive green fluorescence (Figure 4B–D). In contrast, no green fluorescence was detected in placentas developed from uninjected control blastocysts (12 out of 12) (Figure 4E–G), demonstrating the chimerization of TS-like cells. Notably, we did not detect any fluorescence in placental blood vessels (Figure 4D, arrow) and the yolk sac (Figure 4H–I). Taken together, these evidences demonstrated the ability of rabbit TS-like cells to differentiate into trophoblastic derivatives in vitro and in vivo.


Generation of trophoblast stem cells from rabbit embryonic stem cells with BMP4.

Tan T, Tang X, Zhang J, Niu Y, Chen H, Li B, Wei Q, Ji W - PLoS ONE (2011)

Rabbit TS-like cells can chimerize placentae in vivo.(A) GFP transgenic TS-like cells. (B) Whole-mount phase-contrast photographs of the undersides of chimeric placenta. White rectangle represents the part of placenta magnified in (C). (C) Magnification of part of the chimeric placenta in (B). (D) Fluorescent photograph of (C). The placenta showed an extensive contribution of GFP+ cells; Note that the blood vessels of the placenta had no fluorescence (arrow). (E) Whole-mount phase-contrast photographs of the undersides of control placenta developed from un-injected blastocyst. White rectangle represents the part of placenta magnified in (F). (F) Magnification of part of a non-chimeric placenta in (E). (G) Fluorescent photograph of (F). The control placenta showed weak background fluorescence. (H) Phase-contrast photograph of the Yolk sac from the chimeric embryo. (I) Fluorescent photograph of (H) Magnification of C, I, J is 4×, and F is 2×; Magnification of D, E is 7×, and G and H is 4×.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3040765&req=5

pone-0017124-g004: Rabbit TS-like cells can chimerize placentae in vivo.(A) GFP transgenic TS-like cells. (B) Whole-mount phase-contrast photographs of the undersides of chimeric placenta. White rectangle represents the part of placenta magnified in (C). (C) Magnification of part of the chimeric placenta in (B). (D) Fluorescent photograph of (C). The placenta showed an extensive contribution of GFP+ cells; Note that the blood vessels of the placenta had no fluorescence (arrow). (E) Whole-mount phase-contrast photographs of the undersides of control placenta developed from un-injected blastocyst. White rectangle represents the part of placenta magnified in (F). (F) Magnification of part of a non-chimeric placenta in (E). (G) Fluorescent photograph of (F). The control placenta showed weak background fluorescence. (H) Phase-contrast photograph of the Yolk sac from the chimeric embryo. (I) Fluorescent photograph of (H) Magnification of C, I, J is 4×, and F is 2×; Magnification of D, E is 7×, and G and H is 4×.
Mentions: The above evidences prompted us to take the strictest test as if these TS-like cells were able to chimerize the placental tissues when injected into blastocysts. GFP transgenic TS-like cells (Figure 4A) were injected into blastocysts to examine their ability to chimerize the placental tissues. At day 20 of gestation, placenta proper, but not embryo proper of the conceptus (4 out of 12) derived from the green TS like-cell injected blastocysts emitted intensive green fluorescence (Figure 4B–D). In contrast, no green fluorescence was detected in placentas developed from uninjected control blastocysts (12 out of 12) (Figure 4E–G), demonstrating the chimerization of TS-like cells. Notably, we did not detect any fluorescence in placental blood vessels (Figure 4D, arrow) and the yolk sac (Figure 4H–I). Taken together, these evidences demonstrated the ability of rabbit TS-like cells to differentiate into trophoblastic derivatives in vitro and in vivo.

Bottom Line: Trophoblast stem (TS) cells are ideal models to investigate trophectoderm differentiation and placental development.Rabbit ES cells generated in our laboratory were induced to differentiate in the presence of BMP4 and TS-like cell colonies were isolated and expanded.These cells expressed the molecular markers of mouse TS cells, were able to invade, give rise to derivatives of TS cells, and chimerize placental tissues when injected into blastocysts.

View Article: PubMed Central - PubMed

Affiliation: Kunming Primate Research Center, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan, China.

ABSTRACT
Trophoblast stem (TS) cells are ideal models to investigate trophectoderm differentiation and placental development. Herein, we describe the derivation of rabbit trophoblast stem cells from embryonic stem (ES) cells. Rabbit ES cells generated in our laboratory were induced to differentiate in the presence of BMP4 and TS-like cell colonies were isolated and expanded. These cells expressed the molecular markers of mouse TS cells, were able to invade, give rise to derivatives of TS cells, and chimerize placental tissues when injected into blastocysts. The rabbit TS-like cells maintained self-renewal in culture medium with serum but without growth factors or feeder cells, whilst their proliferation and identity were compromised by inhibitors of FGFs and TGF-β receptors. Taken together, our study demonstrated the derivation of rabbit TS cells and suggested the essential roles of FGF and TGF-β signalings in maintenance of rabbit TS cell self-renewal.

Show MeSH
Related in: MedlinePlus